桔小实蝇等五种实蝇的SSR鉴定

Microsatellite Markers DNA for Identification of Four Species of Bactrocera and Bactrocera dorsalis(Hendel) in Chongqing

桔小实蝇Bactrocera(Bactrocera)dorsalis(Hendel)是一种为害多种水果、瓜类、茄类蔬菜的检疫性害虫,食性杂、寄主范围广泛,直接威胁水果和蔬菜的安全生产。采用改进的CTAB法提取单头实蝇的基因组DNA,从20对引物中筛选出2对重复性好、多态性高的引物,运用微卫星标记(SSR)技术,初步构建了桔小实蝇和重庆市常见南亚果实蝇B.(Zeugodacus)tau(Walker)、宽带果实蝇B.a(Z.)scutellata(Hendel)、瓜实蝇B.(Z.)cucurbitae(Coquillett)、黑颜果实蝇B.(Z.)diaphora(Hendel)5种实蝇的指纹图谱。通过这些图谱可容易区分重庆市常见4种实蝇和桔小实蝇,实现了采用2对引物1次扩增的简单快速有效方法将桔小实蝇与其他4种实蝇区分开。这为重庆市桔小实蝇非疫区的建设提供了快速、准确的鉴定方法。

The fruits flies of the Bactrocera （Diptera： Tephritidae） are one of the major economically important insects in Asia. So far, there are at least four species found in Chongqing, China. By using mierosatellite markers （ SSR）, the fingerprint of five Bactrocera species, B. tau （Walker）, B. scutellata （Hendel）, B. cucurbitae （Coquinett）, B. diaphora （Hendel）and B. dorsalis （Hendel）were investigated. C, enomie DNA was extracted from thoracic tissue of individual adult flies by CTAB method and amplified with the polymerase chain reaction. With good repredueibility and high polymorphie ,three from 20 primers were chose to construct primarily the fingerprint. It is easy to distinguish between B. dorsalis and the others through the fingerprint assess that was a simple,quicker and efficient method only applying 2 primers and once amplification. The technique is able to identify fleetly and exactly B. dorsalis for construction pest and diseases free areas of citrus in Chongqing.